[Subspecies identification for Mycobacterium abscessus group].
To identify the subspecies of Mycobacterium abscessus (M.abscessus) group. The corresponding genes (hsp65 and rpoB) in 19 clinical isolates (CIs) of M.abscessus group susceptible to clarithromycin were amplified by PCR. Phylogenetic analyses and subspecies identification of the hsp65 gene and rpoB gene were conducted separately and jointly by using the Mega program and PhyML program, and the conflicting species were further identified by the internally transcribed spacer (ITS) sequencing. The rrl and erm(41) of M.abscessus group detection were performed by PCR sequencing, and MICs of inducible resistance to clarithromycin were determined by the broth microdilution method, and then the phenotypic patterns of 19 isolates were analyzed. Comparisons between hsp65 and rpoB sequences of the 19 clinical isolates led to the identification of 10 CIs as Mycobacterium (M). massiliense, 4 CIs as Mycobacterium (M). abscessus, and 1 CI as Mycobacterium (M). bolletii, while the other 4 isolates were identified as M. massiliense by hsp65 gene sequencing and as M.abscessus by rpoB gene sequencing. Ultimately the 4 conflicting isolates were identified as M. massiliense by ITS sequencing. No mutations in the rrl gene with clarithromycin resistance were found. The -35 sequence of the erm (41) promoter of M.abscessus was different from that of M. bolletii and M. massiliense, and the nucleotide at position 28 was polymorphic (T28 or C28); -35 sequence and the nucleotide at position 28 were the same in erm (41) for M. bolletii and M. massiliense. Fourteen M. massiliense strains shared 100% (14/14) homology for erm (41) with 276 bp deletions and 2 bp deletions, and no deletions were found in 4 CIs as M.abscessus and 1 CIs as M. bolletii. The clarithromycin inducible resistance test showed that 3 M.abscessus with T28 (CI02, CI04, CI12) and 1 M. bolletii (CI18) strains were highly resistant, and the other 14 M. massiliense and 1 M.abscessus with polymorphic C28 (CI17) strains remained susceptible. No correlations between -35 sequence of erm (41) promoter and clarithromycin inducible resistance were found. Hsp65 is applicable to the identification of the subspecies of M.abscessus group, and due to the fact that subspecies of M.abscessus group shows distinct genotytpcally feature in erm (41) sequencing and phenotypic feature in clarithromycin susceptibility, hsp65 and erm (41) can be applied to the identification of subspecies of M.abscessus group, and the resulting data may be useful for clinical diagnosis and treatments.


Management of onchocerciasis. Personal observations.
During a 12-month period (1977 - 1978) of employment in the river forest area of the Ivory Coast, an isolated area hyperendemic for onchocerciasis, a unique opportunity arose to study the disease in depth. Personal observations and experiences with the disease did not necessarily correlate with textbook statements. Although onchocerciasis may involve almost any area of the body it more commonly involves the skin, eyes, lymphatics and groin area. The disease does not appear to cause premature or direct death, but is often acutely and chronically disabling. Diethylcarbamazine and suramin are effective in preventing disabling complications and in diminishing or controlling the progression of complications, but are nephrotoxic. Surgery is utilized to excise microfilaria-producing nodules in order to minimize or control disease progression and for the repair of the frequently occurring complication of inguinal or femoral hernias and hydroceles. A high percentage of patients had microfilarial eye infiltrates but blindness was uncommon. The disease should therefore be known as 'river eye disease' rather than the current 'river blindness'. There is no known method of effective prevention of the disease that is acceptable on a long-term basis.


From benchmarking HITS-CLIP peak detection programs to a new method for identification of miRNA-binding sites from Ago2-CLIP data.
Experimental evidence indicates that about 60% of miRNA-binding activity does not follow the canonical rule about the seed matching between miRNA and target mRNAs, but rather a non-canonical miRNA targeting activity outside the seed or with a seed-like motifs. Here, we propose a new unbiased method to identify canonical and non-canonical miRNA-binding sites from peaks identified by Ago2 Cross-Linked ImmunoPrecipitation associated to high-throughput sequencing (CLIP-seq). Since the quality of peaks is of pivotal importance for the final output of the proposed method, we provide a comprehensive benchmarking of four peak detection programs, namely CIMS, PIPE-CLIP, Piranha and Pyicoclip, on four publicly available Ago2-HITS-CLIP datasets and one unpublished in-house Ago2-dataset in stem cells. We measured the sensitivity, the specificity and the position accuracy toward miRNA binding sites identification, and the agreement with TargetScan. Secondly, we developed a new pipeline, called miRBShunter, to identify canonical and non-canonical miRNA-binding sites based on de novo motif identification from Ago2 peaks and prediction of miRNA::RNA heteroduplexes. miRBShunter was tested and experimentally validated on the in-house Ago2-dataset and on an Ago2-PAR-CLIP dataset in human stem cells. Overall, we provide guidelines to choose a suitable peak detection program and a new method for miRNA-target identification.


Enkephalin peptides in fetal sheep, changes with gestation, origin and production by the placenta.
Enkephalin-containing peptides have been followed in the circulation of fetal sheep between 118-143 days gestation. Using a combination of radioimmunoassay and hplc met5-enkephalin was found in the concentration range 60-500 pg/ml and proenkephalins containing met5-enkephalin had a concentration of 150-4000 pg/ml. The concentration of both increased towards term. The sources of the enkephalin peptides was investigated by measurement of differences across the umbilical circulation and by studying the effects of fetal adrenal demedullation and chemical sympathectomy. The placenta showed a continuous net output of enkephalin peptides which increased close to term. This placental output was increased sharply by reduction of uterine blood flow either using compression of the uterine artery or through infusion of adrenaline at 35 micrograms/min into the maternal circulation. Maternal hypoxia caused by breathing 9% O2 plus 3% CO2 also increased fetal plasma enkephalin levels, although not output from the placenta. Adrenal demedullation, particularly if accompanied by chemical sympathectomy depressed fetal plasma enkephalin concentrations and sharply suppressed the fetal peptide responses to maternal hypoxia. It is concluded that the placenta and the fetal adrenal are important sources of met5-enkephalin-containing peptides in the fetal circulation. The placental production appears to be closely tied to changes in uterine perfusion and adrenal output changes in response to fetal oxygenation.


[Anesthesia in preterm labor].
The authors analyze early neonatal morbidity in 87 preterm babies and the values of perinatal mortality of newborns born to mothers whose labor was anesthesized by different methods. The task of this study was to assess the effect of long epidural anesthesia (LEA) used for preterm labor pain relief on the survival of small-for-date babies. Standard anesthesia of labor is compared to LEA. LEA ensured adequate analgesia and stable hemodynamics; moreover, it protected the intrauterine fetus from hypoxic injuries to the central nervous system due to normalization of uterine contractility and uteroplacental bloodflow and relaxation of the muscles of the fundus of the uterus.


Supramodality effects in visual and haptic spatial processes.
In this article, the authors investigated unimodal and cross-modal processes in spatial working memory. A number of locations had to be memorized within visual or haptic matrices according to different experimental conditions known to be critical in accounting for the effects of perception on imagery. Results reveal that some characteristics of the generated mental image remained strictly inherent to the modality in which information was acquired; in general, accuracy was higher when configurations were visually rather than haptically explored (Experiments 1 and 3). Interestingly, the same pattern emerged when the effects of simultaneous versus sequential processing of the stimuli inherent to vision and haptics were isolated from perceptual modality (Experiment 2). Supramodal elements were also identified (Experiment 3) that were specifically associated to the nature of the cognitive processes, regardless of the original characteristics of the sensory information. These data indicate that both unimodal modality-specific and higher order supramodal mechanisms are simultaneously used in spatial processes.


Measuring what matters to patients: Using goal content to inform measure choice and development.
Personalised care requires personalised outcomes and ways of feeding back clinically useful information to clinicians and practitioners, but it is not clear how to best personalise outcome measurement and feedback using existing standardised outcome measures. The constant comparison method of grounded theory was used to compare goal themes derived from goals set at the outset of therapy for 180 children aged between 4 and 17 years, visiting eight child and adolescent mental health services, to existing standardised outcome measures used as part of common national datasets. In all, 20 out of 27 goal themes corresponded to items on at least one commonly used outcome measure. Consideration of goal themes helped to identify potential relevant outcome measures. However, there were several goal themes that were not captured by items on standardised outcome measures. These seemed to be related to existential factors such as understanding, thinking about oneself and future planning. This presents a powerful framework for how clinicians can use goals to help select a standardised outcome measure (where this is helpful) in addition to the use of a goal-based outcome measure and personalise choices. There may be areas not captured by standardised outcome measures that may be important for children and young people and which may only be currently captured in goal measurement. There is an indication that we may not be measuring what is important to children and young people. We may need to develop or look for new measures that capture these areas.


Quantification of posterior capsule tightness and motion loss in patients with shoulder impingement.
The relationship between posterior capsule tightness and dysfunction has long been recognized clinically but has not been biometrically quantified. The purpose of this study was to quantify changes in range of motion and posterior capsule tightness in patients with dominant or nondominant shoulder impingement. Measurements of posterior capsule tightness and external and internal rotation range of motion were made in 31 patients with shoulder impingement and in 33 controls without shoulder abnormality. Patients with impingement in the nondominant arm had increased posterior capsule tightness and decreased internal and external rotation range of motion compared with controls. Patients with impingement in their dominant arm had increased posterior capsule tightness and reduced internal rotation range of motion but no significant loss of external rotation range of motion compared with controls. Posterior capsule tightness in impingement patients showed a significant correlation with loss of internal rotation range of motion. Patients with shoulder impingement in their nondominant arm had a more global loss of range of motion compared with patients having impingement in their dominant arm. We believe we have described a valid clinical measurement for identifying posterior capsule tightness in patients with shoulder impingement.


Non-superiority of Kakkonto, a Japanese herbal medicine, to a representative multiple cold medicine with respect to anti-aggravation effects on the common cold: a randomized controlled trial.
Kakkonto, a Japanese herbal medicine, is frequently used to treat the common cold not only with a physician's prescription, but also in self-medication situations. This study aimed to examine whether Kakkonto prevents the aggravation of cold symptoms if taken at an early stage of illness compared with a well-selected Western-style multiple cold medicine. This study was a multicenter, active drug-controlled, randomized trial. Adults 18 to 65 years of age who felt a touch of cold symptoms and visited 15 outpatient healthcare facilities within 48 hours of symptoms onset were enrolled. The participants were randomly assigned to two groups: one treated with Kakkonto (Kakkonto Extract-A, 6 g/day) (n=209) and one treated with a Western-style multiple cold medicine (Pabron Gold-A, 3.6 g/day) (n=198) for at most four days. The primary outcome of this study was the aggravation of cold, nasal, throat or bronchial symptoms, scored as moderate or severe and lasting for at least two days within five days after entry into the study. Among the 410 enrollees, 340 (168 in the Kakkonto group and 172 in the Pabron group) were included in the analyses. The proportion of participants whose colds were aggravated was 22.6% in the Kakkonto group and 25.0% in the Pabron group (p=0.66). The overall severity of the cold symptoms was not significantly different between the groups. No harmful adverse events occurred in either group. Kakkonto did not significantly prevent the progression of cold symptoms, even when prescribed at an early stage of the disease.


Hepatitis C virus infection: molecular pathways to metabolic syndrome.
Chronic infection with hepatitis C virus (HCV) can induce insulin resistance (IR) in a genotype-dependent fashion, thus contributing to steatosis, progression of fibrosis and resistance to interferon therapy. The molecular mechanisms in genotype 1 patients that lead to metabolic syndrome are still ambiguous. Based on our current understanding, HCV proteins associate with mitochondria and endoplasmic reticulum and promote oxidative stress. The latter mediates signals involving the p38 mitogen-activated protein kinase and activates nuclear factor kappa B. This transcription factor plays a key role in the expression of cytokines, tumor necrosis factor alpha (TNF-alpha), interleukin 6, interleukin 8, tumor growth factor beta, and Fas ligand. TNF-alpha inhibits the function of insulin receptor substrates and decreases the expression of the glucose transporter and lipoprotein lipase in peripheral tissues, which is responsible for the promotion of insulin resistance. Furthermore, reduced adiponectin levels, loss of adiponectin receptors, and decreased anti-inflammatory peroxisome proliferator-activated receptor alpha in the liver of HCV patients may contribute to reduced fatty acid oxidation, inflammation, and eventually lipotoxicity. This chain of events may be initiated by HCV-associated IR and provides a direction for future research in the areas of therapeutic intervention.


LeafScope: A Portable High-Resolution Multispectral Imager for In Vivo Imaging Soybean Leaf.
Portable devices for measuring plant physiological features with their isolated measuring chamber are playing an increasingly important role in plant phenotyping. However, currently available commercial devices of this type, such as soil plant analysis development (SPAD) meter and spectrometer, are dot meters that only measure a small region of the leaf, which does not perfectly represent the highly varied leaf surface. This study developed a portable and high-resolution multispectral imager (named LeafScope) to in-vivo image a whole leaf of dicotyledon plants while blocking the ambient light. The hardware system is comprised of a monochrome camera, an imaging chamber, a lightbox with different bands of light-emitting diodes (LEDs) array, and a microcontroller. During measuring, the device presses the leaf to lay it flat in the imaging chamber and acquires multiple images while alternating the LED bands within seconds in a certain order. The results of an experiment with soybean plants clearly showed the effect of nitrogen and water treatments as well as the genotype differences by the color and morphological features from image processing. We conclude that the low cost and easy to use LeafScope can provide promising imaging quality for dicotyledon plants, so it has great potential to be used in plant phenotyping.


Principles of Clinical Ethics and Their Application to Practice.
An overview of ethics and clinical ethics is presented in this review. The 4 main ethical principles, that is beneficence, nonmaleficence, autonomy, and justice, are defined and explained. Informed consent, truth-telling, and confidentiality spring from the principle of autonomy, and each of them is discussed. In patient care situations, not infrequently, there are conflicts between ethical principles (especially between beneficence and autonomy). A four-pronged systematic approach to ethical problem-solving and several illustrative cases of conflicts are presented. Comments following the cases highlight the ethical principles involved and clarify the resolution of these conflicts. A model for patient care, with caring as its central element, that integrates ethical aspects (intertwined with professionalism) with clinical and technical expertise desired of a physician is illustrated.


Septic phlebitis caused by Candida albicans and diagnosed by needle aspiration.
We have described a patient who had Candida albicans suppurative thrombophlebitis at the site of an indwelling intravenous catheter. The microbial etiology was established by needle aspiration of the venipuncture site, affording rapid and accurate identification of the infecting organism and providing prompt institution of therapy. Phlebectomy and a brief course of amphotericin B prevented further dissemination of the infection.


The Fe/S ratio of pyrrhotite group sulfides in chondrites: An indicator of oxidation and implications for return samples from asteroids Ryugu and Bennu.
Determining compositional trends among individual minerals is key to understanding the thermodynamic conditions under which they formed and altered, and is also essential to maximizing the scientific value of small extraterrestrial samples, including returned samples and meteorites. Here we report the chemical compositions of Fe-sulfides, focusing on the pyrrhotite-group sulfides, which are ubiquitous in chondrites and are sensitive indicators of formation and alteration conditions in the protoplanetary disk and in small Solar System bodies. Our data show that while there are trends with the at.% Fe/S ratio of pyrrhotite with thermal and aqueous alteration in some meteorite groups, there is a universal trend between the Fe/S ratio and degree of oxidation. Relatively reducing conditions led to the formation of troilite during: (1) chondrule formation in the protoplanetary disk (i.e., pristine chondrites) and (2) parent body thermal alteration (i.e., LL4 to LL6, CR1, CM, and CY chondrites). Oxidizing and sulfidizing conditions led to the formation of Fe-depleted pyrrhotite with low Fe/S ratios during: (1) aqueous alteration (i.e., CM and CI chondrites), and (2) thermal alteration (i.e., CK and R chondrites). The presence of troilite in highly aqueously altered carbonaceous chondrites (e.g., CY, CR1, and some CM chondrites) indicates they were heated after aqueous alteration. The presence of troilite, Fe-depleted pyrrhotite, or pyrite in a chondrite can provide an estimate of the oxygen and sulfur fugacities at which it was formed or altered. The data reported here can be used to estimate the oxygen fugacity of formation and potentially the aqueous and/or thermal histories of sulfides in extraterrestrial samples, including those returned by the Hayabusa2 mission and due to be returned by the OSIRIS-REx mission in the near future.


Transthoracic approach for Pott's disease.
Spinal tuberculosis with paraplegia, although decreasing in incidence, remains a problem in certain sections of the United States and in most underdeveloped nations. Evacuations of the tuberculous abscess, debridement of necrotic bone, and fusion of the anterior spine are maneuvers performed increasingly by thoracic surgeons. Twenty-two patients with Pott's disease and symptoms of back pain, gibbous deformity, and neurological deficit underwent thoracotomy. Nine were paraplegic prior to operation. Postoperatively, spinal fusion occurred in all cases. All paraplegic patients can walk now, and 17 of the 22 treated operatively have been completely rehabilitated and are in school or working. The average hospital stay was 2.4 months. There was 1 operative death. Medical treatment for Pott's disease consisted of bed rest and chemotherapy and resulted in progressive neurological deficit in 2 patients with an average hospital stay of 2.2 years. We believe operative treatment offers two distinct advantages: excellent reversal of even long-standing paraplegia and markedly shortened hospital stay.


Relationship between nurses' organizational trust levels and their organizational citizenship behaviors.
This research used a descriptive and explorative design to determine the levels of nurses' organizational trust and organizational citizenship and to investigate relationships between the levels of organizational trust and organizational citizenship behaviors. Nurses who had completed their orientation from a total of 11 hospitals with bed capacities of 100 and located in the European district of Istanbul were included in the sample for this study. Formal, written applications and approval of the ethical committee were obtained from concerned institutions before proceeding with the data collection step. The Organizational Trust Inventory and the Organizational Citizenship Level Scale, a questionnaire form including five questions regarding nurses' personal characteristics, were used in data collection. Data collection tools were distributed to 900 nurses in total, and usable data were obtained from 482 nurses. Number and percentage calculations and Pearson correlation analysis were used to assess research data. The results of the present research showed that nurses had a higher than average level of trust in their managers and coworkers and they trusted more in their managers and coworkers than their institutions. The Organizational Citizenship Level Scale indicated that the behavior most frequently demonstrated by the nurses was conscientiousness, followed by courtesy and civic virtue, whereas sportsmanship was displayed to an average extent. An analysis of relationships between nurses' level of organizational trust and their organizational citizenship behaviors revealed that nurses who trust in their managers, institutions, and coworkers demonstrated the organizational citizenship behaviors of conscientiousness, civic virtue, courtesy, and altruism more frequently. The findings attained in this study indicated that the organizational trust the staff had in their institutions, managers, and coworkers influenced the organizational citizenship behaviors of conscientiousness, civic virtue, altruism, and courtesy, whereas it had no effect on sportsmanship behavior. Nurse managers should introduce studies to improve their subordinates' organizational trust to ensure that they develop organizational citizenship behaviors, and they should support them in this process. These topics for nursing services will provide guidance to managers, particularly to managers of nursing services, in establishing processes to predict nurses' organizational commitment, job satisfaction, performance, intention to leave, and other relevant issues.


Height loss in older women: risk of hip fracture and mortality independent of vertebral fractures.
We examined if height loss in older women predicts risk of hip fractures, other nonspine fractures, and mortality, and whether this risk is independent of both vertebral fractures (VFx) and bone mineral density (BMD) by dual-energy X-ray absorptiometry. Among 3124 women age 65 and older in the Study of Osteoporotic Fractures, we assessed the association with measured height change between year 0 (1986-1988) and year 15 (2002-2004) and subsequent risk of radiologically confirmed hip fractures, other nonspine fractures, and mortality assessed via death certificates. Follow-up occurred every 4 months for fractures and vital status (>95% contacts complete). Cox proportional hazards models assessed risk of hip fracture, nonspine fracture, and mortality over a mean of 5 years after height change was assessed (ie, after final height measurement). After adjustment for VFx, BMD, and other potential covariates, height loss >5 cm was associated with a marked increased risk of hip fracture [hazard ratio (HR) 1.50, 95% confidence interval (CI) 1.06, 2.12], nonspine fracture (HR 1.48; 95% CI 1.20, 1.83), and mortality (1.45; 95% CI 1.21, 1.73). Although primary analyses were a subset of 3124 survivors healthy enough to return for a year 15 height measurement, a sensitivity analysis in the entire cohort (n = 9677) using initial height in earlier adulthood [self-reported height at age 25 (-40 years) to measured height age >65 years (Year 0)] demonstrated consistent results. Height loss >5 cm (2″) in older women was associated with a nearly 50% increased risk of hip fracture, nonspine fracture, and mortality-independent of incident VFx and BMD.


The Integrator complex regulates differential snRNA processing and fate of adult stem cells in the highly regenerative planarian Schmidtea mediterranea.
In multicellular organisms, cell type diversity and fate depend on specific sets of transcript isoforms generated by post-transcriptional RNA processing. Here, we used Schmidtea mediterranea, a flatworm with extraordinary regenerative abilities and a large pool of adult stem cells, as an in vivo model to study the role of Uridyl-rich small nuclear RNAs (UsnRNAs), which participate in multiple RNA processing reactions including splicing, in stem cell regulation. We characterized the planarian UsnRNA repertoire, identified stem cell-enriched variants and obtained strong evidence for an increased rate of UsnRNA 3'-processing in stem cells compared to their differentiated counterparts. Consistently, components of the Integrator complex showed stem cell-enriched expression and their depletion by RNAi disrupted UsnRNA processing resulting in global changes of splicing patterns and reduced processing of histone mRNAs. Interestingly, loss of Integrator complex function disrupted both stem cell maintenance and regeneration of tissues. Our data show that the function of the Integrator complex in UsnRNA 3'-processing is conserved in planarians and essential for maintaining their stem cell pool. We propose that cell type-specific modulation of UsnRNA composition and maturation contributes to in vivo cell fate choices, such as stem cell self-renewal in planarians.


Factors affecting the course and severity of transnasally induced Staphylococcus aureus pneumonia in mice.
In order to examine several factors that may affect the course and severity of transnasally induced Staphylococcus aureus pneumonia in mice, bacteria were prepared in a free suspension or bound to fetal mouse cells. Immunosuppression was induced in five strains of mice (ICR, C57BL/6, BALB/c, C3H/He and CBA/J) by injection of cyclophosphamide (200 mg/kg body weight), 2 days before infection. Impairment of mucociliary clearance was induced by intranasal instillation of formalin. Mice were then infected with various doses and strains of the organism. Although no significant differences were observed between either form of inoculum, pretreatment with formalin plus cyclophosphamide was associated with a significant increase in lung bacterial counts. In particular, cyclophosphamide treatment was associated with a high mortality in mice infected with several strains of S. aureus irrespective of their toxin production profiles. Histopathological examination demonstrated that in mice treated with formalin plus cyclophosphamide, clusters of bacteria were observed in lung parenchyma, associated with a mild accumulation of inflammatory cells at day 2 and extensive cell infiltration at day 7. CBA/J mice represented the most susceptible strain among those examined, with 10(4)- and 10(2)-fold higher bacterial counts in the lungs at days 3 and 5, respectively. These results indicate that neutropenia and impaired mucociliary clearance are major factors that influence the severity of S. aureus pneumonia in mice. Analysis of the role of genetic background in enhancement of vulnerability to infection is warranted in future studies.


Prevalence and genetic characterization of shiga toxin-producing Escherichia coli isolates from slaughtered animals in Bangladesh.
To determine the prevalence of Shiga toxin (Stx)-producing Escherichia coli (STEC) in slaughter animals in Dhaka, Bangladesh, we collected rectal contents immediately after animals were slaughtered. Of the samples collected from buffalo (n = 174), cows (n = 139), and goats (n = 110), 82.2%, 72.7%, and 11.8% tested positive for stx(1) and/or stx(2), respectively. STEC could be isolated from 37.9%, 20.1%, and 10.0% of the buffalo, cows, and goats, respectively. STEC O157 samples were isolated from 14.4% of the buffalo, 7.2% of the cows, and 9.1% of the goats. More than 93% (n = 42) of the STEC O157 isolates were positive for the stx(2), eae, katP, etpD, and enterohemorrhagic E. coli hly (hly(EHEC)) virulence genes. STEC O157 isolates were characterized by seven recognized phage types, of which types 14 (24.4%) and 31 (24.4%) were predominant. Subtyping of the 45 STEC O157 isolates by pulsed-field gel electrophoresis showed 37 distinct restriction patterns, suggesting a heterogeneous clonal diversity. In addition to STEC O157, 71 STEC non-O157 strains were isolated from 60 stx-positive samples from 23.6% of the buffalo, 12.9% of the cows, and 0.9% of the goats. The STEC non-O157 isolates belonged to 36 different O groups and 52 O:H serotypes. Unlike STEC O157, most of the STEC non-O157 isolates (78.9%) were positive for stx(1). Only 7.0% (n = 5) of the isolates were positive for hly(EHEC), and none was positive for eae, katP, and etpD. None of the isolates was positive for the iha, toxB, and efa1 putative adhesion genes. However, 35.2% (n = 25), 11.3% (n = 8), 12.7% (n = 9), and 12.7% (n = 9) of the isolates were positive for the lpf(O113), saa, lpfA(O157/01-141), and lpfA(O157/OI-154) genes, respectively. The results of this study provide the first evidence that slaughtered animals like buffalo, cows, and goats in Bangladesh are reservoirs for STEC, including the potentially virulent STEC strain O157.


Differential pH Dynamics in Synaptic Vesicles From Intact Glutamatergic and GABAergic Synapses.
Synaptic transmission requires the presynaptic release of neurotransmitter from synaptic vesicles (SVs) onto the postsynaptic neuron. Vesicular neurotransmitter transporter proteins, which use a V-ATPase-generated proton gradient, play a crucial role in packaging neurotransmitter into SVs. Recent work has revealed different proton dynamics in SVs expressing the vesicular glutamate transporter (VGLUT) or the vesicular GABA transporter (VGAT) proteins. At the whole synapse level, this results in different steady-state pH and different reacidification dynamics during SV recycling (Egashira et al., 2016). In isolated SVs, the presence of VGAT causes a higher steady state pH, which is correlated with a faster proton efflux rate (Farsi et al., 2016). To address whether proton efflux from GABAergic and glutamatergic SVs in intact synapses differs, we applied a glutamatergic- or GABAergic neuron-specific expression strategy (Chang et al., 2014) to express a genetically encoded pH sensor (synaptophysin pHluorin; SypHy) and/or light-activated proton pump (pHoenix; (Rost et al., 2015). We confirm, with SypHy post-stimulation fluorescence dynamics, that the pH profile of recycling GABAergic SVs differs from that of recycling glutamatergic SVs (Egashira et al., 2016). Using light-activation of pHoenix in pH-neutral vesicles, we investigated the pH dynamics of actively filling vesicles, and could show that proton efflux from GABAergic SVs is indeed initially faster than glutamatergic SVs in intact synapses. Finally, we compared the filling rate of empty glutamatergic and GABAergic vesicles using pHoenix as a proton source, and find a slightly faster filling of glutamatergic vs. GABAergic SVs.


Acetaminophen-Induced Rat Hepatotoxicity Based on M1/M2-Macrophage Polarization, in Possible Relation to Damage-Associated Molecular Patterns and Autophagy.
Overdose of acetaminophen (APAP), an antipyretic drug, is an important cause of liver injury. However, the mechanism in the rat model remains undetermined. We analyzed APAP-induced hepatotoxicity using rats based on M1/M2-macrophage functions in relation to damage-associated molecular patterns (DAMPs) and autophagy. Liver samples from six-week-old rats injected with APAP (1000 mg/kg BW, ip, once) after 15 h fasting were collected at hour 10, and on days 1, 2, 3, and 5. Liver lesions consisting of coagulation necrosis and inflammation were seen in the affected centrilobular area on days 1 and 2, and then, recovered with reparative fibrosis by day 5. Liver exudative enzymes increased transiently on day 1. CD68+ M1-macrophages increased significantly on days 1 and 2 with increased mRNAs of M1-related cytokines such as IFN-g and TNF-α, whereas CD163+ M2-macrophages appeared later on days 2 and 3. Macrophages reacting to MHC class II and Iba1 showed M1-type polarization, and CD204+ macrophages tended to be polarized toward M2-type. At hour 10, interestingly, HMGB1 (representative DAMPs) and its related signals, TLR-9 and MyD88, as well as LC3B+ autophagosomes began to increase. Collectively, the pathogenesis of rat APAP hepatotoxicity, which is the first, detailed report for a rat model, might be influenced by macrophage functions of M1 type for tissue injury/inflammation and M2-type for anti-inflammatory/fibrosis; particularly, M1-type may function in relation to DAMPs and autophagy. Understanding the interplayed mechanisms would provide new insight into hepato-pathogenesis and contribute to the possible development of therapeutic strategies.


Combination Therapy of High-flow Nasal Cannula and Upper-body Elevation for Postoperative Sleep-disordered Breathing: Randomized Crossover Trial.
The low acceptance rate of continuous positive airway pressure therapy in postoperative patients with untreated obstructive sleep apnea (OSA) indicates the necessity for development of an alternative postoperative airway management strategy. The authors considered whether the combination of high-flow nasal cannula and upper-body elevation could improve postoperative OSA. This nonblinded randomized crossover study performed at a single university hospital investigated the effect on a modified apnea hypopnea index, based exclusively on the airflow signal without arterial oxygen saturation criteria (flow-based apnea hypopnea index, primary outcome), of high-flow nasal cannula (20 l · min-1 with 40% oxygen concentration) with and without upper-body elevation in patients with moderate to severe OSA. Preoperative sleep studies were performed at home (control, no head-of-bed elevation) and in hospital (30-degree head-of-bed elevation). On the first and second postoperative nights, high-flow nasal cannula was applied with or without 30-degree head-of-bed elevation, assigned in random order to 23 eligible participants. Twenty-two of the 23 (96%) accepted high-flow nasal cannula. Four participants resigned from the study. Control flow-based apnea hypopnea index (mean ± SD, 60 ± 12 events · h-1; n = 19) was reduced by 15 (95% CI, 6 to 30) events · h-1 with head-of-bed elevation alone (P = 0.002), 10.9 (95% CI, 1 to 21) events · h-1 with high-flow nasal cannula alone (P = 0.028), and 23 (95% CI, 13 to 32) events · h-1 with combined head-of-bed elevation and high-flow nasal cannula (P < 0.001). Compared to sole high-flow nasal cannula, additional intervention with head-of-bed elevation significantly decreased flow-based apnea hypopnea index by 12 events · h-1 (95% CI, 2 to 21; P = 0.022). High-flow nasal cannula, alone or in combination with head-of-bed elevation, also improved overnight oxygenation. No harmful events were observed. The combination of high-flow nasal cannula and upper-body elevation reduced OSA severity and nocturnal hypoxemia, suggesting a role for it as an alternate postoperative airway management strategy.


Vulnerable exons, like ACADM exon 5, are highly dependent on maintaining a correct balance between splicing enhancers and silencers.
It is now widely accepted that aberrant splicing of constitutive exons is often caused by mutations affecting cis-acting splicing regulatory elements, but there is a misconception that all exons have an equal dependency on splicing regulatory elements and thus a similar susceptibility to aberrant splicing. We investigated exonic mutations in ACADM exon 5 to experimentally examine their effect on splicing and found that 7 out of 11 tested mutations affected exon inclusion, demonstrating that this constitutive exon is particularly vulnerable to exonic splicing mutations. Employing ACADM exon 5 and 6 as models, we demonstrate that the balance between splicing enhancers and silencers, flanking intron length, and flanking splice site strength are important factors that determine exon definition and splicing efficiency of the exon in question. Our study shows that two constitutive exons in ACADM have different inherent vulnerabilities to exonic splicing mutations. This suggests that in silico prediction of potential pathogenic effects on splicing from exonic mutations may be improved by also considering the inherent vulnerability of the exon. Moreover, we show that single nucleotide polymorphism that affect either of two different exonic splicing silencers, located far apart in exon 5, all protect against both immediately flanking and more distant exonic splicing mutations.


[Condyloma acuminatum: detection of capsid antigen of human papillomavirus and microscopic study].
420 cases of condyloma acuminata and condyloma-like lesions in vulvar region in Guangzhou area were studies on paraffin sections: detection of capsid antigen of human papilloma-versus (HPV-Ag) and observation of microscopic morphology. Of the 420 cases, 160 cases (38.1%) were diagnosed condyloma acuminata. In which, diagnostic koilocytes can be found in 156 cases (97.5%) and HPV-Ag were positive in 105 cases (65.6%). 260 cases (61.9%) were diagnosed chronic inflammation of the skin or mucosa and were all negative for HPV-Ag, koilocyte-like cells but diagnostic koilocytes presented in these lesions. In this paper, morphological features of diagnostic koilocytes and koilocyte-like cells were detailed, and histological and gross distinction between condyloma and condyloma-like lesions were described.


Differential expression of thrombospondin and cellular fibronectin during remodeling in proliferative glomerulonephritis.
Thrombospondin-1 (TSP-1) and an alternatively spliced fibronectin (Fn)-EIIIA isoform are adhesive proteins associated with embryogenesis and tissue remodeling. We compared, by immunohistochemistry and in situ hybridization, the course of TSP-1 and Fn-EIIIA expression in a model of glomerulonephritis induced by Habu snake venom (HV) and characterized by mesangial cell migration, proliferation, and extracellular matrix (ECM) synthesis. At 24 hr after HV, TSP-1 and Fn-EIIIA proteins localized in the central aspects of lesions associated with platelets and macrophages and at the margins of lesions coinciding with mesangial cell migration (determined by Thy-1 staining). Mesangial cells at this time expressed TSP-1 but not Fn-EIIIA mRNA. TSP-1 protein and mRNA peaked in lesions at 48 hr and were associated with cell proliferation (determined by PCNA, alpha-smooth muscle actin phenotype, and expression of beta-PDGF receptor mRNA). TSP-1 expression declined at 72 hr when expression of ECM synthesis peaked, as determined by increased expression of collagen Type IV, laminin, and TGF-beta1 protein and mRNA. Mesangial cell expression of Fn-EIIIA was first observed at 48 hr and was most abundant at 72 hr after HV. Therefore, platelet- and macrophage-derived Fn-EIIIA and TSP-1 in early lesions are associated with mesangial cell migration. Mesangial cell upregulation of TSP-1 is associated with migration and proliferation but not maximal ECM accumulation, whereas mesangial cell expression of Fn-EIIIA is associated with proliferation and ECM accumulation. These results suggest distinctive temporal and spatial roles for TSP-1 and Fn-EIIIA in remodeling during glomerular disease. (J Histochem Cytochem 47:533-543, 1999)


Work-practice changes associated with an electronic emergency department whiteboard.
Electronic whiteboards are introduced at emergency departments (EDs) to improve work practices. This study investigates whether the time physicians and nurses at an ED spend in patient rooms versus at the control desk increases after the introduction of an electronic whiteboard. After using this whiteboard for four months nurses, but not physicians, spend more of their time with the patients. With the electronic whiteboard, nurses spend 28% of their time in patient rooms and physicians 20%. Importantly, the changes facilitated by the electronic whiteboard are also dependent on implementation issues, existing work practices and the clinicians' experience. Another change in the work practices is distributed access to whiteboard information from the computers in patient rooms. A decrease in the mental workload of the coordinating nurse was envisaged but has not emerged. Achieving more changes appears to require an increase in whiteboard functionality and a firmer grip on the implementation process.


Study of the lactation curve in dairy cattle on farms in central Mexico.
Accurate knowledge of lactation curves has an important relevance to management and research of dairy production systems. A number of equations have been proposed to describe the lactation curve, the most widely applied being the gamma equation. The objective of this work was to compare and evaluate candidate functions for their predictive ability in describing lactation curves from central Mexican dairy cows reared under 2 contrasting management systems. Five equations were considered: Gaines (exponential decay), Wood (gamma equation), Rook (Michaelis-Menten xexponential), and 2 more mechanistic ones (Dijkstra and Pollott). A database consisting of 701 and 1283 records of cows in small-scale and intensive systems, respectively, was used in the analysis. Before analysis, the database was divided into 6 groups representing first, second, and third and higher parity cows in both systems. In all cases except second and above parity cows in small-scale systems, all models improved on the Gaines equation. The Wood equation explained much of the variation, but its parameters do not have direct biological interpretation. Although the Rook equation fitted the data well, some of the parameter estimates were not significant. The Dijkstra equation consistently gave better predictions, and its parameters were usually statistically significant and lend themselves to physiological interpretation. As such, the differences between systems and parity could be explained due to variations in theoretical initial milk production at parturition, specific rates of secretory cell proliferation and death, and rate of decay, all of which are parameters in the model. The Pollott equation, although containing the most biology, was found to be over-parameterized and resulted in nonsignificant parameter estimates. For central Mexican dairy cows, the Dijkstra equation was the best option to use in describing the lactation curve.


Neonatal abstinence syndrome.
Neonatal abstinence syndrome (NAS) is a result of the sudden discontinuation of fetal exposure to substances that were used or abused by the mother during pregnancy. Withdrawal from licit or illicit substances is becoming more common among neonates in both developed and developing countries. NAS continues to be an important clinical entity throughout much of the world. NAS leads to a constellation of signs and symptoms involving multiple systems. The pathophysiology of NAS is not completely understood. Urine or meconium confirmation may assist the diagnosis and management of NAS. The Finnegan scoring system is commonly used to assess the severity of NAS; scoring can be helpful for initiating, monitoring, and terminating treatment in neonates. Nonpharmacological care is the initial treatment option, and pharmacological treatment is required if an improvement is not observed after nonpharmacological measures or if the infant develops severe withdrawal. Morphine is the most commonly used drug in the treatment of NAS secondary to opioids. An algorithmic approach to the management of infants with NAS is suggested. Breastfeeding is not contraindicated in NAS, unless the mother is taking street drugs, is involved in polydrug abuse, or is infected with HIV. Future studies are required to assess the long-term effects of NAS on children after prenatal exposure.


The MOBI-Kids Study Protocol: Challenges in Assessing Childhood and Adolescent Exposure to Electromagnetic Fields from Wireless Telecommunication Technologies and Possible Association with Brain Tumor Risk.
The rapid increase in mobile phone use in young people has generated concern about possible health effects of exposure to radiofrequency (RF) and extremely low frequency (ELF) electromagnetic fields (EMF). MOBI-Kids, a multinational case-control study, investigates the potential effects of childhood and adolescent exposure to EMF from mobile communications technologies on brain tumor risk in 14 countries. The study, which aims to include approximately 1,000 brain tumor cases aged 10-24 years and two individually matched controls for each case, follows a common protocol and builds upon the methodological experience of the INTERPHONE study. The design and conduct of a study on EMF exposure and brain tumor risk in young people in a large number of countries is complex and poses methodological challenges. This manuscript discusses the design of MOBI-Kids and describes the challenges and approaches chosen to address them, including: (1) the choice of controls operated for suspected appendicitis, to reduce potential selection bias related to low response rates among population controls; (2) investigating a young study population spanning a relatively wide age range; (3) conducting a large, multinational epidemiological study, while adhering to increasingly stricter ethics requirements; (4) investigating a rare and potentially fatal disease; and (5) assessing exposure to EMF from communication technologies. Our experience in thus far developing and implementing the study protocol indicates that MOBI-Kids is feasible and will generate results that will contribute to the understanding of potential brain tumor risks associated with use of mobile phones and other wireless communications technologies among young people.


Changes in predator exposure, but not in diet, induce phenotypic plasticity in scorpion venom.
Animals embedded between trophic levels must simultaneously balance pressures to deter predators and acquire resources. Venomous animals may use venom toxins to mediate both pressures, and thus changes in this balance may alter the composition of venoms. Basic theory suggests that greater exposure to a predator should induce a larger proportion of defensive venom components relative to offensive venom components, while increases in arms races with prey will elicit the reverse. Alternatively, reducing the need for venom expenditure for food acquisition, for example because of an increase in scavenging, may reduce the production of offensive venom components. Here, we investigated changes in scorpion venom composition using a mesocosm experiment where we manipulated scorpions' exposure to a surrogate vertebrate predator and live and dead prey. After six weeks, scorpions exposed to surrogate predators exhibited significantly different venom chemistry compared with naive scorpions. This change included a relative increase in some compounds toxic to vertebrate cells and a relative decrease in some compounds effective against their invertebrate prey. Our findings provide, to our knowledge, the first evidence for adaptive plasticity in venom composition. These changes in venom composition may increase the stability of food webs involving venomous animals.


The contribution of upper limb and total body movement to adolescents' energy expenditure whilst playing Nintendo Wii.
Little research documents the contribution of upper limb and total body movement to energy expenditure (EE) during active video gaming. To address this, EE, heart rate (HR), and, upper limb and total body movement were assessed in 11- to 17-year-old adolescents whilst playing three active (Nintendo Wii) and one sedentary (XBOX 360) video games. Non-dominant upper limb activity, EE and HR were significantly greater during Wii Sports boxing [mean 267.2 (SD 115.8) J kg(-1) min(-1); 136.7 (24.5) beats min(-1)] than tennis or bowling (P < or = 0.044). For all active games hip activity best predicted EE (R (2) > or = 0.53), with two-measure models of HR and single-site activity data, and multi-site activity data, similarly explaining the variance in EE (R (2) > or = 0.64). The physiological cost of upper-body orientated active video games increased when movement of both upper limbs was encouraged. Improvements in EE explanatory power provide support for multi-site activity monitoring during unique, non-ambulatory activities.


Herpes simplex virus induces the replication of foreign DNA.
Plasmids containing the simian virus 40 (SV40) DNA replication origin and the large T gene are replicated efficiently in Vero monkey cells but not in rabbit skin cells. Efficient replication of the plasmids was observed in rabbit skin cells infected with herpes simplex virus type 1 (HSV-1) and HSV-2. The HSV-induced replication required the large T antigen and the SV40 replication origin. However, it produced concatemeric molecules resembling replicative intermediates of HSV DNA and was sensitive to phosphonoacetate at concentrations known to inhibit the HSV DNA polymerase. Therefore, it involved the HSV DNA polymerase itself or a viral gene product(s) which was expressed following the replication of HSV DNA. Analyses of test plasmids lacking SV40 or HSV DNA sequences showed that, under some conditions, HSV also induced low-level replication of test plasmids containing no known eucaryotic replication origins. Together, these results show that HSV induces a DNA replicative activity which amplifies foreign DNA. The relevance of these findings to the putative transforming potential of HSV is discussed.


Characteristics of pediatric patients seen in medical school-based osteopathic manipulative medicine clinics.
Manual medicine--specifically osteopathic manipulative treatment (OMT)--is commonly used in treating patients aged 18 years or younger. However, no published reports have described characteristics of this patient population or the conditions for which OMT is used with these patients. To better counsel parents, train physicians and other healthcare providers, and prioritize research, an improved understanding of the use of OMT in children is needed. To characterize pediatric patients and their conditions as seen in a medical school-based osteopathic manipulative medicine clinic. Retrospective analysis of administrative data on the use of OMT. Faculty osteopathic manipulative medicine specialty clinics associated with the University of New England College of Osteopathic Medicine. Data were analyzed from patients seen in the clinics from January 1, 2007, through December 31, 2007, if they were younger than 19 years at their first visit during that period. Factors included in the data analysis were patient age at first visit, age at time of visit, number of visits during the study period, types of clinical diagnoses, and visits with nonmusculoskeletal diagnoses. A total of 407 patients generated 1500 clinic visits. Data showed a mean of 3.7 visits per patient (25th-75th percentiles = 2-5 visits) during the 1-year study period. The mean age at the first clinic visit was 7 years, 3 months, with the 25th-to-75th percentile being 1 year, 9 months, to 12 years, 3 months. Clinic visits by age group (ie, age at time of visit) as percentages of total visits were as follows: 0 to 11 months, 13.7%; 1 to 4 years, 33.3%; 5 to 12 years, 28.9%, older than 12 years, 24.2%. Diagnoses provided in visits covered a wide variety of common pediatric conditions. For the entire study population, 43.5% of visits included nonmusculoskeletal diagnoses. The percentages of visits with nonmusculoskeletal diagnoses for each age group were as follows: 0 to 11 months, 33.7%; 1 to 4 years, 64.0%; 5 to 12 years, 48.8%; older than 12 years, 17.7%. Pediatric patients seen in the faculty osteopathic manipulative medicine specialty clinic included the entire pediatric age range and a wide range of common pediatric conditions. A substantial number of visits involved treatment for nonmusculoskeletal conditions. Further investigation is needed to determine if the patient sample of the present study is representative of other clinical settings or geographic regions.


Clinical investigation of retinitis pigmentosa.
The diagnostic features and future research directions of retinitis pigmentosa were documented in this update and review of the subject. An extensive and current bibliography is provided.


[The value of radiological follow-up for stage III melanoma].
The modalities of follow-up (frequency of consultations and interest of repeated radiological examinations) of patients presenting with glandular metastases of melanoma (stage III of the AJCC classification) have not reached a consensus. Since 1995, we have proposed clinical follow-up every two months and radiological controls with a thoracic-abdominal-pelvic scan every 4 months, to patients at high risk of relapse for the early screening of an infra-clinical relapse. The median follow-up was of 16 months (range: 1 to 82 months). Eight patients out of 24 (33 p. 100) followed-up in this manner, had asymptomatic metastases discovered by the radiological examinations. Among these 8 patients, three presented with a an operable, single, metastatic localization and two patients underwent surgery. One patient relapsed 3 months later, the other was still alive without relapse 24 months later. Surgery remains the treatment of choice for all stages of melanoma. In the absence of clearly effective treatment of metastatic melanoma, the early discovery of an infra-clinical metastatic relapse presents two major advantages. The first is the discovery of a single, operable metastasis, as was the case in two of the patients out of 24. The second is to be able to suspend an eventual adjuvant therapy with interferon alpha, as soon as a relapse has been discovered.


The requirement for nucleoporin NUP153 during human immunodeficiency virus type 1 infection is determined by the viral capsid.
Lentiviruses likely infect nondividing cells by commandeering host nuclear transport factors to facilitate the passage of their preintegration complexes (PICs) through nuclear pore complexes (NPCs) within nuclear envelopes. Genome-wide small interfering RNA screens previously identified karyopherin β transportin-3 (TNPO3) and NPC component nucleoporin 153 (NUP153) as being important for infection by human immunodeficiency virus type 1 (HIV-1). The knockdown of either protein significantly inhibited HIV-1 infectivity, while infection by the gammaretrovirus Moloney murine leukemia virus (MLV) was unaffected. Here, we establish that primate lentiviruses are particularly sensitive to NUP153 knockdown and investigate HIV-1-encoded elements that contribute to this dependency. Mutants lacking functional Vpr or the central DNA flap remained sensitive to NUP153 depletion, while MLV/HIV-1 chimera viruses carrying MLV matrix, capsid, or integrase became less sensitive when the latter two elements were substituted. Two capsid missense mutant viruses, N74D and P90A, were largely insensitive to NUP153 depletion, as was wild-type HIV-1 when cyclophilin A was depleted simultaneously or when infection was conducted in the presence of cyclosporine A. The codepletion of NUP153 and TNPO3 yielded synergistic effects that outweighed those calculated based on individual knockdowns, indicating potential interdependent roles for these factors during HIV-1 infection. Quantitative PCR revealed normal levels of late reverse transcripts, a moderate reduction of 2-long terminal repeat (2-LTR) circles, and a relatively large reduction in integrated proviruses upon NUP153 knockdown. These results suggest that capsid, likely by the qualities of its uncoating, determines whether HIV-1 requires cellular NUP153 for PIC nuclear import.


Revealing shape selectivity and catalytic activity trends within the pores of H-ZSM-5 crystals by time- and space-resolved optical and fluorescence microspectroscopy.
A combination of in-situ optical and fluorescence microspectroscopy has been employed to investigate the oligomerization of styrene derivatives occurring in the micropores of coffin-shaped H-ZSM-5 zeolite crystals in a space- and time-resolved manner. The carbocationic intermediates in this reaction act as reporter molecules for catalytic activity, since they exhibit strong optical absorption and fluorescence. In this way, reactant selectivity and restricted transition-state selectivity for 14 substituted styrene molecules can be visualized and quantified. Based on a thorough analysis of the time- and space-resolved UV/Vis spectra, it has been revealed that two main parameters affect the reaction rates, namely, the carbocation stabilization effect and the diffusion hindrance. The stabilization effect was tested by comparison of the reaction rates for 4-methoxystyrene versus 4-methylstyrene and in the series 4-bromo-, 4-chloro and 4-fluorostyrene; in both cases less electronegative substituents were found to accelerate the reaction. As to the steric effect, bulkier chemical groups bring down the reaction rate, as evident from the observation that 4-methoxystyrene is more reactive than 4-ethoxystyrene due to differences in their diffusivity, while heavily substituted styrenes, such as 3,4-dichlorostyrene and 2,3,4,5,6-pentafluorostyrene, cannot enter the zeolite pore system and therefore do not display any reactivity. Furthermore, beta-methoxystyrene and trans-beta-methylstyrene show limited reactivity as well as restricted reaction-product formation due to steric constraints imposed by the H-ZSM-5 channel system. Finally, polarized-light optical microspectroscopy and fluorescence microscopy demonstrate that dimeric styrene compounds are predominantly formed and aligned within the straight channels at the edges of the crystals, whereas a large fraction of trimeric carbocations along with dimeric compounds are present in the straight channels of the main body of the H-ZSM-5 crystals. Our results reinforce the observation of a non-uniform catalytic behavior within zeolite crystals, with specific parts of the zeolite grains being less accessible and reactive towards reactant molecules. The prospects and potential of this combined in-situ approach for studying large zeolite crystals in the act will be discussed.


Validation of the unruptured intracranial aneurysm treatment score: comparison with real-world cerebrovascular practice.
OBJECTIVE The purpose of this study was to compare the unruptured intracranial aneurysm treatment score (UIATS) recommendations with the real-world experience in a quaternary academic medical center with a high volume of patients with unruptured intracranial aneurysms (UIAs). METHODS All patients with UIAs evaluated during a 3-year period were included. All factors included in the UIATS were abstracted, and patients were scored using the UIATS. Patients were categorized in a contingency table assessing UIATS recommendation versus real-world treatment decision. The authors calculated the percentage of misclassification, sensitivity, specificity, and area under the receiver operating characteristic (ROC) curve. RESULTS A total of 221 consecutive patients with UIAs met the inclusion criteria: 69 (31%) patients underwent treatment and 152 (69%) did not. Fifty-nine (27%) patients had a UIATS between -2 and 2, which does not offer a treatment recommendation, leaving 162 (73%) patients with a UIATS treatment recommendation. The UIATS was significantly associated with treatment (p < 0.001); however, the sensitivity, specificity, and percentage of misclassification were 49%, 80%, and 28%, respectively. Notably, 51% of patients for whom treatment would be recommended by the UIATS did not undergo treatment in the real-world cohort and 20% of patients for whom conservative management would be recommended by UIATS had intervention. The area under the ROC curve was 0.646. CONCLUSIONS Compared with the authors' experience, the UIATS recommended overtreatment of UIAs. Although the UIATS could be used as a screening tool, individualized treatment recommendations based on consultation with a cerebrovascular specialist are necessary. Further validation with longitudinal data on rupture rates of UIAs is needed before widespread use.


Comparative analysis of genetic modifiers in Drosophila points to common and distinct mechanisms of pathogenesis among polyglutamine diseases.
Spinocerebellar Ataxia type 1 (SCA1) and Huntington's disease (HD) are two polyglutamine disorders caused by expansion of a CAG repeat within the coding regions of the Ataxin-1 and Huntingtin proteins, respectively. While protein folding and turnover have been implicated in polyglutamine disorders in general, many clinical and pathological differences suggest that there are also disease-specific mechanisms. Taking advantage of a collection of genetic modifiers of expanded Ataxin-1-induced neurotoxicity, we performed a comparative analysis in Drosophila models of the two diseases. We show that while some modifier genes function similarly in SCA1 and HD Drosophila models, others have model-specific effects. Surprisingly, certain modifier genes modify SCA1 and HD models in opposite directions, i.e. they behave as suppressors in one case and enhancers in the other. Furthermore, we find that modulation of toxicity does not correlate with alterations in the formation of neuronal intranuclear inclusions. Our results point to potential common therapeutic targets in novel pathways, and to genes and pathways responsible for differences between Ataxin-1 and Huntingtin-induced neurodegeneration.


Visual Aesthetic Perceptions and Preferences in Conserved Objects of Plain Silk: Comparison of Three Conservation Methods.
Three conservation methods were executed on bonnets in plain monochrome silk, to investigate which method is perceived as the most visually aesthetic; 11 bonnets were produced, 10 given identical damages, and 9 were conserved, 3 with each method. The damage was secured onto a support fabric with laid couching, a long stitch fastened with short perpendicular stitches, or brick couching, short stitches placed like brick-work, or covered with crepeline (a semi-transparent silk). The participants were 30 Swedish textile conservators (29 women; ages 29-78 years, M = 51.9, SD = 12.9), and 30 museum visitors (20 women; ages 15-74 years, M = 41.1, SD = 18.3). The participants' task was to rate the bonnets on a 100-point continuous preference scale, based on how visually attractive they found each bonnet. Preferences were compared between the two groups of participants and the conservation methods. The bonnets with crepeline were the most preferred, and those with laid couching were the least preferred, among both groups of participants.


Screening for Chronic Obstructive Pulmonary Disease (COPD) in an Urban HIV Clinic: A Pilot Study.
Increased smoking and a detrimental response to tobacco smoke in the lungs of HIV/AIDS patients result in an increased risk for COPD. We aimed to determine the predictive value of a COPD screening strategy validated in the general population and to identify HIV-related factors associated with decreased lung function. Subjects at least 35 years of age at an HIV clinic in New York City completed a COPD screening questionnaire and peak flow measurement. Those with abnormal results and a random one-third of normal screens had spirometry. 235 individuals were included and 89 completed spirometry. Eleven (12%) had undiagnosed airway obstruction and 5 had COPD. A combination of a positive questionnaire and abnormal peak flow yielded a sensitivity of 20% (specificity 93%) for detection of COPD. Peak flow alone had a sensitivity of 80% (specificity 80%). Abnormal peak flow was associated with an AIDS diagnosis (p=0.04), lower nadir (p=0.001), and current CD4 counts (p=0.001). Nadir CD4 remained associated in multivariate analysis (p=0.05). Decreased FEV1 (<80% predicted) was associated with lower CD4 count nadir (p=0.04) and detectable current HIV viral load (p=0.01) in multivariate analysis. Questionnaire and peak flow together had low sensitivity, but abnormal peak flow shows potential as a screening tool for COPD in HIV/AIDS. These data suggest that lung function may be influenced by HIV-related factors.


[Apolipoprotein E genotype as a risk factor in Japanese patients with early-onset and late-onset Alzheimer's disease].
Recent studies have provided evidence of an association of apolipoprotein E (apoE) epsilon 4 allele and late-onset sporadic Alzheimer's disease (AD). Some studies have shown the possibility that apoE epsilon 4 is a risk factor of developing AD in early-onset type. We have analyzed the apoE gene polymorphism in a sample of 310 Japanese AD subjects and 237 age-matched Japanese controls. We divided the sporadic AD patients into two subgroups of 237 late-onset (> 65 years) and 73 early-onset (< or = 65 years) patients, and into three subgroups according to their apoE genotype, no epsilon 4, one epsilon 4, and two epsilon 4 alleles. Our data confirmed an association between epsilon 4 allele and early-onset AD and late-onset AD. The odds ratios (95% confidence interval) referred to no epsilon 4 allele for AD were 3.4 (1.7-7.0) for one epsilon 4 allele and 20.3 (2.5-166.6) for two epsilon 4 alleles in early-onset type, and 6.7 (3.9-11.3) for one epsilon 4 allele and 19.0 (2.5-145.6) for two epsilon 4 alleles in late-onset type. These ratios were significantly increased in both early-onset AD and late-onset AD. Kaplan-Meier survival analysis, which estimates the age of onset for subjects with no, one, and two epsilon 4 alleles in early-onset and late-onset type, revealed a significant dose effect where each additional epsilon 4 allele made the age of onset earlier (p < 0.0001). The age of onset is 9.7 years earlier for two epsilon 4 bearers and 3.9 years earlier for one epsilon 4 bearers than no epsilon 4 bearers in late-onset AD, 2.9 years earlier for two epsilon 4 bearers and 1.4 years earlier for one epsilon 4 bearers than no epsilon 4 bearers in early-onset AD. Moreover, we studied an association between apoE epsilon 2 allele and early-onset AD and late-onset AD. There was a significantly decreased frequency of apoE epsilon 2 allele in patients with late-onset AD (p = 0.026), although the frequency of apoE epsilon 2 was not changed significantly in early-onset AD (p = 0.360). The odds ratios referred to no epsilon 2 allele for AD were 1.9 (0.6-5.7) for one epsilon 2 allele in early-onset type, and 0.4 (0.2-0.9) for one epsilon 2 allele in late-onset type. Our study suggested the difference in the effect of apoE genotype on developing AD between early-onset and late-onset type in Japanese patients.


Response of vitamin B-6 deficiency and the carpal tunnel syndrome to pyridoxine.
The specific activities and percentage deficiencies of the glutamic oxaloacetic transaminase of erythrocytes (EGOT) were determined for patients with carpal tunnel syndrome (CTS) diagnosed by clinical examination and electrical conduction data; the EGOT data revealed a severe deficiency of vitamin B-6. After double-blind treatment with pyridoxine and placebo, two physicians identified those receiving pyridoxine (clinically improved) and those receiving placebo (did not improve) without error, P less than 0.0078. Correcting a deficiency of the coenzyme at receptors of existing molecules of the apoenzyme appears to take place within days; correction of the deficiency in the number of molecules of the transaminase takes place over 10-12 weeks. The clinical response, appraised by the diminution of the symptoms of CTS, was correlated only with the restored levels of the transaminase which presumably results from a translational long-term increase in the number of molecules of EGOT by a mechanism activated by correcting a deficiency of pyridoxal 5'-phosphate. Apparent Km values of EGOT were identical for groups of patients with CTS and others without CTS but with identical specific activities, indicating that CTS is a primary deficiency of vitamin B-6 rather than one of a dependency state. Clinical improvement of the syndrome with pyridoxine therapy may frequently obviate hand surgery.


Single-Molecule Imaging Demonstrates Ligand Regulation of the Oligomeric Status of CXCR4 in Living Cells.
The role of dimerization and oligomerization of G-protein-coupled receptors in their signal transduction is highly controversial. Delineating this issue can greatly facilitate rational drug design. With single-molecule imaging, we show that chemokine receptor CXCR4 exists mainly as a monomer in normal mammalian living cells and forms dimers and higher-order oligomers at a high expression level, such as in cancer cells. Chemotaxis tests demonstrate that the signal transduction activity of CXCR4 does not depend only on its expression level, indicating a close relation with the oligomeric status of CXCR4. Moreover, binding ligands can effectively upregulate or downregulate the oligomeric level of CXCR4, which suggests that binding ligands may realize their pivotal roles by regulating the oligomeric status of CXCR4 rather than by simply inducing conformational changes.


Swept Source-Optical Coherence Tomography Angiography for Management of Secondary Choroidal Neovascularization in Punctate Inner Choroidopathy.
The purpose was to demonstrate the diagnostic and therapeutic feasibility of swept source-optical coherence tomography angiography (SS-OCTA) by picturing neovascular changes secondary to a rare white dot syndrome following long-term intravitreal ranibizumab (IVR). A 28-year-old Caucasian myopic female presented with visual loss in her right eye only. The clinical examination and multimodal imaging including spectral domain (SD)-OCT, blue-peak autofluorescence, fluorescein, and indocyanine green angiography (HRA Spectralis, Heidelberg Engineering; Heidelberg, Germany) as well as SS-OCTA (DRI Triton, Topcon; Tokyo, Japan) led to the diagnosis of idiopathic punctate inner choroidopathy with secondary subfoveal choroidal neovascularization (CNV). In addition to oral corticosteroids, a pro re nata regimen with IVR was initiated and guided by repeated SD-OCT and SS-OCTA. Six IVR were administered based on functional SS-OCTA en face scans illustrating vessel transformation and downsizing of the CNV area while SD-OCT B-scans were inconclusive as indirect signs of activity were absent throughout the follow-up period. SS-OCTA provided new possibilities for monitoring vessel development. IVR was managed based on vessel density as displayed by SS-OCTA.


2-[N-(X-chlorophenyl)carbamoyl]benzenesulfonamide (with X = 2 and 4).
The structures of 2-[N-(2-chlorophenyl)carbamoyl]benzenesulfonamide and 2-[N-(4-chlorophenyl)carbamoyl]benzenesulfonamide, both C(13)H(11)ClN(2)O(3)S, are stabilized by extensive intra- and intermolecular hydrogen bonds. In both structures, sulfonamide groups are hydrogen bonded via the N and O atoms and form chains of molecules. The carbamoyl groups are also hydrogen bonded, involving the O and N atoms, further strengthening the polymeric chains running along the c and a axes in the 2- and 4-chloro derivatives, respectively. Carbamoylsulfonamide derivatives are novel compounds with a great potential for medicinal applications.


Alterations of the cyclin D1/p16-pRB pathway in mantle cell lymphoma.
Mantle cell lymphoma (MCL) has recently become generally accepted as a subentity of malignant lymphomas that is characterized by the chromosomal translocation t(11;14)(q13;q32), resulting in the overexpression of cyclin D1. Cyclin D1 forms a complex with cell cycle-dependent kinase (cdk) 4, which inactivates the retinoblastoma protein (pRB) via phosphorylation. However, in transgenic mice, the overexpression of cyclin D1 alone is not sufficient for the development of malignant lymphoma. To determine whether other members of the pRB pathway contribute to the malignant transformation of MCL, we analyzed 37 cases of MCL that were well characterized by morphology, immunophenotype, and/or interphase cytogenetics [detection of t(11;14)(q13;q32)]. Interphase fluorescence in situ hybridization was performed using a cosmid contig (250 kb) of the CDKN2/p16 region (encoding an inhibitor of the cyclin D1/cdk4 complex) and a phage contig (200 kb) of the Rb region. CDKN2/p16 deletion was detected in 15 cases (41%), including 6 homozygous deletions; Rb was deleted in 15 cases (41%), all of which were hemizygous deletions. Nine cases (24%) had deletions of both CDKN2/p16 and Rb. Further analysis of a subset of 17 MCLs revealed a highly significant correlation between CDKN2/p16 deletion and proliferation index, determined by the rate of Ki67 expression (P = 0.014; t test). No significant correlation was found between CDKN2/p16 deletion and the blastoid variant of MCL (P = 0.23; Fisher's test) or between proliferation index and blastoid morphology (P = 0.51; t test). Deletion of Rb did not have any impact on cell proliferation in addition to CDKN2/p16 deletion (P = 0.76; t test). Additional analysis of 13q14 deletions suggests that these deletions may target another gene telomeric to Rb. We conclude that deletion of CDKN2/p16 occurs in approximately one-half of MCLs and is a more relevant indicator of the proliferative features as compared to morphological criteria. In contrast, although deletions of chromosomal band 13q14 are frequent in MCL, inactivation of Rb seems not to be involved in the pathogenesis of MCL.


Anti-Biofouling and Self-Cleaning Surfaces Featured with Magnetic Artificial Cilia.
The fouling of surfaces submerged in a liquid is a serious problem for many applications including lab-on-a-chip devices and marine sensors. Inspired by the versatility of cilia in manipulating fluids and particles, it is experimentally demonstrated that surfaces partially covered with magnetic artificial cilia (MAC) have the capacity to efficiently prevent attachment and adhesion of real biofouling agents-microalgae Scenedesmus sp. Actuation of the MAC resulted in over 99% removal of the algae for two different scenarios: (1) actuating the MAC immediately after injecting the algae into a microfluidic chip, demonstrating antifouling and (2) starting to actuate the MAC 1 week after injecting the algae into the chip and leaving them to grow in static conditions, showing self-cleaning. It is shown that the local and global flows generated by the actuated MAC are substantial, resulting in hydrodynamic shear forces acting on the algae, which are likely to be key to efficient antifouling and self-cleaning. These findings and insights will potentially lead to novel types of self-cleaning and antifouling strategies, which may have a relevant practical impact on different fields and applications including lab-on-a-chip devices and water quality analyzers.


Surveillance approaches among colorectal cancer survivors after curative-intent.
Intensive surveillance among colorectal cancer patients receiving curative-intent, particularly during the first 2 to 3 years of follow-up, has a beneficial impact upon all-cause survival at five years. Intensive surveillance appears to be associated with the early detection of recurrences, and more often accompanied by a clinical presentation enabling surgical resection. The optimal combination and frequency of surveillance tests is unknown. Imaging of the chest and abdomen have increasingly been recommended by professional organizations, in addition to CEA levels, in order to detect resectable recurrences. Metachronous cancers are relatively uncommon, nonetheless, surveillance colonoscopy is typically recommended. Cau-tion is warranted in further advancing the intensity of surveillance, as increasingly aggressive surveillance programs risk increased detection of pseudodisease. Different types of information will enhance our understanding of the impact of follow-up programs, including data regarding quality of life, cost, and patient preferences.


Development of a palladium-catalyzed decarboxylative cross-coupling of (2-azaaryl)carboxylates with aryl halides.
A catalytic method for the decarboxylative coupling of 2-(azaaryl)carboxylates with aryl halides is described. The decarboxylative cross-coupling presented is mediated by a system catalytic in both palladium and copper without requiring stoichiometric amounts of organometallic reagents or organoboronic acids. This method circumvents additional synthetic steps required to prepare 2-azaaryl organometallics and organoborates as nucleophilic coupling partners, which are prone to protodemetallation and protodeborylation and produce potentially toxic byproducts.


Recent Advances of Wearable Antennas in Materials, Fabrication Methods, Designs, and Their Applications: State-of-the-Art.
The demand for wearable technologies has grown tremendously in recent years. Wearable antennas are used for various applications, in many cases within the context of wireless body area networks (WBAN). In WBAN, the presence of the human body poses a significant challenge to the wearable antennas. Specifically, such requirements are required to be considered on a priority basis in the wearable antennas, such as structural deformation, precision, and accuracy in fabrication methods and their size. Various researchers are active in this field and, accordingly, some significant progress has been achieved recently. This article attempts to critically review the wearable antennas especially in light of new materials and fabrication methods, and novel designs, such as miniaturized button antennas and miniaturized single and multi-band antennas, and their unique smart applications in WBAN. Finally, the conclusion has been drawn with respect to some future directions.


Brain injury: prolonged induction of transcription factors.
A specific temporal order of events at the cellular and molecular level occurs in response to injury to the brain. Injury-compromised neurons degenerate while surviving neurons undergo neuritogenesis and synaptogenesis to establish neuronal connectivity destroyed in the injury. Several genes, such as those coding cytoskeletal proteins and growth factors, have been shown to be regulated by AP-1 and NF-kappa B transcription factors, two of the most studied DNA binding regulatory proteins. Our laboratory has discovered that Fos-related antigen-2 from AP-1 transcription factor family and NF-kappa B p65 and p50 subunits are induced long-term (days to months) in the brain after neurotoxic, excitotoxic or ischemic insult. Fos-related antigen-2 is induced in neurons in several models of injury and its elevated expression lasts days to months, corresponding to the severity. The time-course of FRA-2 induction is abbreviated with less severe insult (terminal damage) relative to the cell death, but the induction occurs during the period of regeneration and repair in both models. NF-kappa B p65 is basally expressed in hippocampal and cortical neurons, but is elevated in reactive astrocytes in hippocampus and entorhinal cortex starting at two days and lasting at least two weeks after kainate treatment. Neurons of the hippocampus surviving ischemic or neurotoxic injury increase expression of NF-kappa B p50 for at least a week after injury, suggesting a function for p50 in neuronal survival and/or repair. The extended expression of these transcription factors implies a role in the activation of genes related to repair and regeneration, such as growth factors and synaptic proteins, after injury to the CNS.


The nature and role of pigments of marine invertebrates.
Marine animals, especially those from tropical waters, are often brilliantly coloured, and bright colouration is widespread in both sessile and non-sessile invertebrates. These spectacular natural colours are common in species inhabiting shallow waters, and appear not only in animals exposed to bright light, but also in those living in dark areas where colours are visible only with artificial illumination. Marine organisms also show variation in colour with depth and geographical location, and display great variety in colour patterning. These colour characteristics are the result of several different processes, and serve various purposes - the distribution and function of pigments seems to vary between invertebrate groups. In addition to playing an important role in how marine organisms interact, pigments may be involved in physiological processes. Although nitrogenous pigments predominate, marine organisms contain pigments belonging to all the major structural classes of natural products, as well as some that are unique to the marine environment. This review discusses the nature and significance of such pigments, the chemical and biological processes involved, the factors responsible for and affecting bright colourations, as well as their evolution and speculation as to their function.


Sensitivity of anti-Leu-M1 as a marker in Hodgkin's disease.
Seventy-eight cases of Hodgkin's disease (52 nodular sclerosis, 13 mixed cellularity, seven lymphocyte predominance, one lymphocyte depletion, one interfollicular, and four unclassified cases) were stained with anti-Leu-M1 using the immunoperoxidase technique on either formalin- or B-5-fixed, paraffin-embedded sections. The cytoplasmic pattern and extent of staining of Reed-Sternberg cells and mononuclear variants were recorded. Ninety-four percent of the cases had Leu-M1-positive cells. The staining intensity and number of positive cells were greatest in nodular sclerosis and least in lymphocyte predominance. There was variability, however, in the patterns and amounts of positivity for all subtypes. The neoplastic cells in 15 non-Hodgkin's lymphomas were uniformly negative for Leu-M1. For the individual case, lack of Leu-M1 positivity does not exclude Hodgkin's disease as a diagnostic possibility, but when adequate numbers of Reed-Sternberg-like cells are present, complete absence of staining suggests an alternative diagnosis.


Capecitabine for locally advanced and metastatic colorectal cancer: A review.
Capecitabine (Xeloda(®)) is an oral fluoropyrimidine which is produced as a pro-drug of fluorouracil, and shows improved tolerability and intratumor drug concentrations following its tumor-specific conversion to the active drug. We have searched the Pubmed and Cochrane databases from 1980 to 2009 with the purpose of reviewing all available information on Capecitabine, focusing on its clinical effectiveness against colorectal cancer. Special attention has been paid to trials that compared Capecitabine with standard folinic acid (leucovorin, LV)-modulated intravenous 5-fluorouracil (5-FU) bolus regimens in patients with metastatic colorectal cancer. Moreover the efficacy of Capecitabine on metastatic colorectal cancer, either alone or in various combinations with other active drugs such as Irinotecan and Oxaliplatin was also assessed. Finally, neoadjuvant therapy consisting of Capecitabine plus radiation therapy, for locally advanced rectal cancer was analysed. This combination of chemotherapy and radiotherapy has a special role in tumor down staging and in sphincter preservation for lower rectal tumors. Comparative trials have shown that Capecitabine is at least equivalent to the standard LV-5-FU combination in relation to progression-free and overall survival whilst showing a better tolerability profile with a much lower incidence of stomatitis. It is now known that Capecitabine can be combined with other active drugs such as Irinotecan and Oxaliplatin. The combination of Oxaliplatin with Capecitabine represents a new standard of care for metastatic colorectal cancer. Combinating the Capecitabine-Oxaliplatin regimen with promising new biological drugs such as Bevacizumab seems to give a realistic prospect of further improvement in time to progression of metastatic disease. Moreover, preoperative chemo-radiation using oral capecitabine is better tolerated than bolus 5-FU and is more effective in the promotion of both down-staging and sphincter preservation in patients with locally advanced rectal cancer. Finally, the outcomes of recently published trials suggest that capecitabine seems to be more cost effective than other standard treatments for the management of patients with colorectal cancer.


ESI formation of a Meisenheimer complex from tetryl and its unusual dissociation.
The reactivity of the explosive tetryl (N-methyl-N,2,4,6-tetranitroaniline; Mw = 287 u) was studied using electrospray ionization in negative mode. The main species detected in the spectrum corresponds to the ion observed at m/z 318 (previously assumed to be the odd-electron ion [tetryl + HNO](-•), C7H6O9N6). In this study, we show using D-labeling combined with high-resolution mass spectrometry that this species corresponds to an even-electron anion (i.e. C8H8O9N5), resulting from the formation of a Meisenheimer complex between tetryl and the methanol used as the solvent. Fragmentation of this complex under CID conditions revealed an unexpected fragment: the formation of a 2,4,6-trinitrophenoxide anion at m/z 228. (18)O-labeling combined with quantum chemical calculations helped us better understand the reaction pathways and mechanisms involved in the formation of this product ion. This occurs via a transition state leading to a SN2-type reaction, consequently evolving toward an ion-dipole complex. The latter finally dissociates into deprotonated picric acid.


Intraoperative Electrocochleographic Characteristics of Auditory Neuropathy Spectrum Disorder in Cochlear Implant Subjects.
Auditory neuropathy spectrum disorder (ANSD) is characterized by an apparent discrepancy between measures of cochlear and neural function based on auditory brainstem response (ABR) testing. Clinical indicators of ANSD are a present cochlear microphonic (CM) with small or absent wave V. Many identified ANSD patients have speech impairment severe enough that cochlear implantation (CI) is indicated. To better understand the cochleae identified with ANSD that lead to a CI, we performed intraoperative round window electrocochleography (ECochG) to tone bursts in children (n = 167) and adults (n = 163). Magnitudes of the responses to tones of different frequencies were summed to measure the "total response" (ECochG-TR), a metric often dominated by hair cell activity, and auditory nerve activity was estimated visually from the compound action potential (CAP) and auditory nerve neurophonic (ANN) as a ranked "Nerve Score". Subjects identified as ANSD (45 ears in children, 3 in adults) had higher values of ECochG-TR than adult and pediatric subjects also receiving CIs not identified as ANSD. However, nerve scores of the ANSD group were similar to the other cohorts, although dominated by the ANN to low frequencies more than in the non-ANSD groups. To high frequencies, the common morphology of ANSD cases was a large CM and summating potential, and small or absent CAP. Common morphologies in other groups were either only a CM, or a combination of CM and CAP. These results indicate that responses to high frequencies, derived primarily from hair cells, are the main source of the CM used to evaluate ANSD in the clinical setting. However, the clinical tests do not capture the wide range of neural activity seen to low frequency sounds.


Tripartite upstream promoter element essential for expression of Saccharomyces cerevisiae ribosomal protein genes.
To initiate a genetic analysis of yeast ribosomal protein gene promoters, we have constructed a gene fusion between the yeast ribosomal protein gene RP39A and the Escherichia coli lacZ gene. This gene fusion contains approximately 1,030 nucleotides of the 5' flanking region and the first 49 1/3 codons of RP39A fused in frame to a large 3' end fragment of lacZ. Whether it is introduced into yeast cells on a moderately high-copy-number plasmid, or integrated into the yeast genome at the RP39A locus, this RP39A-lacZ gene directs the synthesis of a hybrid transcript which encodes beta-galactosidase activity. Deletions in the 5' flanking region of RP39A-lacZ were constructed by linker insertion and BAL 31 mutagenesis. The expression of the mutant genes in yeast cells was assayed by measuring RP39A-lacZ mRNA and beta-galactosidase levels. By these means we have shown that the sequences between nucleotides -256 and -170 upstream of RP39A are essential for expression of this gene. Three sequence motifs, HOMOL1, RPG, and a T-rich region, which were found in that order 5'----3' upstream of most yeast ribosomal protein genes, were present within this interval. We found that substitution of the CYC1-lacZ upstream activation site with the fragment from nucleotides -298 to -172 upstream of RP39A, containing the HOMOL1-RPG-T-rich motif in that 5'----3' orientation, fully restored expression of the CYC1-lacZ gene. The essentially of HOMOL1, the RPG sequence, and the T-rich region for wild-type levels of expression of RP39A, the conserved location and order of these sequence motifs in yeast ribosomal protein genes, and the ability of a DNA fragment carrying these three sequence elements to substitute for the upstream activation site regions of CYC1 indicate that these three oligonucleotides may be essential to the transcription of yeast ribosomal protein genes.


Structure-property relationships in a series of diglycerol tetraether model lipids and their lyotropic assemblies: the effect of branching topology and chirality.
Three novel diglycerol tetraether lipids with one membrane-spanning chain have been synthesized. These lipids contain only two or four racemic methyl branches at selected positions of the hydrophobic chains in contrast to natural lipids from archaebacterial membranes with an isoprenoid substitution pattern. The insertion of the methyl moieties was realized starting from either (RS)-citronellyl bromide or the inexpensive methyl malonic acid ethyl ester. For chain elongation the Cu-catalysed Grignard coupling reaction was used. The preparation of diglycerol tetraethers was either performed by condensing suitable blocked monoglycerol diethers by Grubbs metathesis or by reaction of the transmembrane C32-chain with blocked glycerols followed by further alkylation steps. Finally, we could show that the resulting lipids can form closed lipid vesicles comparable to the optically pure counterparts. Therefore, these much simpler lipids compared to the natural lipids from archaebacterial membranes are also suitable for preparation of stable tailored liposomes.


Hospital admissions and place of death of residents of care homes receiving specialist healthcare services: A systematic review without meta-analysis.
To synthesize evidence on the ability of specialist care home support services to prevent hospital admission of older care home residents, including at end of life. Systematic review, without meta-analysis, with vote counting based on direction of effect. Fourteen electronic databases were searched from January 2010 to January 2019. Reference lists of identified reviews, study protocols and included documents were scrutinized for further studies. Papers on the provision of specialist care home support that addressed older, long-term care home residents' physical health needs and provided comparative data on hospital admissions were included. Two reviewers undertook study selection and quality appraisal independently. Vote counting by direction of effect and binomial tests determined service effectiveness. Electronic searches identified 79 relevant references. Combined with 19 citations from an earlier review, this gave 98 individual references relating to 92 studies. Most were from the UK (22), USA (22) and Australia (19). Twenty studies were randomized controlled trials and six clinical controlled trials. The review suggested interventions addressing residents' general health needs (p < .001), assessment and management services (p < .0001) and non-training initiatives involving medical staff (p < .0001) can reduce hospital admissions, while there was also promising evidence for services targeting residents at imminent risk of hospital entry or post-hospital discharge and training-only initiatives. End-of-life care services may enable residents to remain in the home at end of life (p < .001), but the high number of weak-rated studies undermined confidence in this result. This review suggests specialist care home support services can reduce hospital admissions. More robust studies of services for residents at end of life are urgently needed. The review addressed the policy imperative to reduce the avoidable hospital admission of older care home residents and provides important evidence to inform service design. The findings are of relevance to commissioners, providers and residents.


Consequences of symbiont co-infections for insect host phenotypes.
Most animals host communities of symbiotic bacteria. In insects, these symbionts may have particularly intimate interactions with their hosts: many are intracellular and can play important roles in host ecology and evolution, including protection against natural enemies. We investigated how interactions between different species or strains of endosymbiotic bacteria within an aphid host influence the outcome of symbiosis for both symbiont and host. We first asked whether different combinations of facultative symbiont species or strains can exist in stable co-infections. We then investigated whether the benefits that facultative bacteria confer on their hosts (protection against natural enemies) are enhanced, reduced or unaltered by the presence of a co-infecting symbiont. We asked this both for co-infecting symbionts that confer different phenotypes on their hosts (protection against fungal pathogens vs. parasitoid wasps) and symbionts with overlapping functions. Finally, we investigated the additional survival costs to aphids of carrying multiple infections of symbiont species or strains, and compared symbiont titres in double and single infections. We found that stable co-infections were possible between all of the combinations of facultative symbiont species (Regiella insecticola + Hamiltonella defensa, Regiella + Rickettsiella sp., Regiella + Spiroplasma sp.) and strains (Hamiltonella) that we studied. Where symbionts provided protection against different natural enemies, no alteration in protection was observed in the presence of co-infections. Where symbionts provided protection against the same natural enemy, the level of protection corresponded to the higher of the two symbionts present. In some instances, aphid hosts suffered additional survival costs when hosting double infections. In the case of Hamiltonella, however, infection with multiple strains of the same symbiont led to lower symbiont titres than single infections, and actually improved aphid survival. We conclude that the long-term maintenance of symbiont co-infections in aphids is likely to be determined primarily by costs of co-infections and in some instances by redundancy of symbiont benefits.


Marden-Walker syndrome: case report, nosologic discussion and aspects of counseling.
The Marden-Walker syndrome is characterized by a mask-like face with blepharophimosis, micrognathia, cleft or high-arched palate, low-set ears, congenital joint contractures, decreased muscular mass, failure to thrive and psychomotor retardation. We report a boy with a phenotype mostly resembling the condition named Marden-Walker syndrome, with many of the criteria proposed for diagnosing this particular phenotype. In addition he had hypoplastic corpus callosum, cerebellar vermis hypoplasia, enlarged cisterna magna and vertebral abnormalities. During pregnancy there were reduced fetal movements. In the present patient the fetal hypokinesia sequence, due to central nervous system malformation, is most compatible with the diagnosis of Marden-Walker syndrome. The etiology is probably heterogeneous, but the possibility of autosomal recessive inheritance should be considered in genetic counseling.


Generation and characterization of a human single-chain fragment variable (scFv) antibody against cytosine deaminase from yeast.
The ability of cytosine deaminase (CD) to convert the antifungal agent 5-fluorocytosine (5-FC) into one of the most potent and largely used anticancer compound such as 5-fluorouracil (5-FU) raised considerable interest in this enzyme to model gene or antibody - directed enzyme-prodrug therapy (GDEPT/ADEPT) aiming to improve the therapeutic ratio (benefit versus toxic side-effects) of cancer chemotherapy. The selection and characterization of a human monoclonal antibody in single chain fragment (scFv) format represents a powerful reagent to allow in in vitro and in vivo detection of CD expression in GDEPT/ADEPT studies. An enzymatic active recombinant CD from yeast (yCD) was expressed in E. coli system and used as antigen for biopanning approach of the large semi-synthetic ETH-2 antibody phage library. Several scFvs were isolated and specificity towards yCD was confirmed by Western blot and ELISA. Further, biochemical and functional investigations demonstrated that the binding of specific scFv with yCD did not interfere with the activity of the enzyme in converting 5-FC into 5-FU. The construction of libraries of recombinant antibody fragments that are displayed on the surface of filamentous phage, and the selection of phage antibodies against target antigens, have become an important biotechnological tool in generating new monoclonal antibodies for research and clinical applications. The scFvH5 generated by this method is the first human antibody which is able to detect yCD in routinary laboratory techniques without interfering with its enzymatic function.